Registration Dossier

Ecotoxicological information

Short-term toxicity to fish

Currently viewing:

Administrative data

Link to relevant study record(s)

Reference
Endpoint:
short-term toxicity to fish
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2017-04-12 to 2017-06-30
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Reference:
Composition 0
Composition 0
Qualifier:
according to
Guideline:
OECD Guideline 203 (Fish, Acute Toxicity Test)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Test material information:
Composition 1
Specific details on test material used for the study:
SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 201601004
- Expiration date of the lot/batch: 01.01.2019

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Store in dark, cool, dry place in tightly closed receptacles
- Stability under test conditions: stable

Analytical monitoring:
yes
Details on sampling:
The samples were filled into 10 mL glass vials and measured at the same day. The sampling was conducted according to the following specification:
- At the start of the test (0 h), treatment NC* and A* were sampled (each group: 3 samples of 6 mL).
- After 24 h, 48 h and 72 h exposure:
- Before renewal of the test solutions, treatment NC* and A* were sampled (each group: 3 samples of 6 mL).
- After renewal of the test solutions, treatment NC* and A* were sampled (each group: 3 samples of 6 mL).
- At the end of the test (96 h), treatment NC* and A* were sampled (each group: 3 samples of 6 mL).

* nominal concentrations (mg/L): 0.0 (Sample code: NC), 5 (Sample code: A)

Chemical analysis of the test item concentration in the test solutions was performed to determine exposure concentration and stability of the test item during exposure. Substance specific analysis was performed at the test site menal GmbH. Of each sampled treatment, one of the samples from 0 h, 24 h, 48 h, 72 h (before and after renewal) and 96 h was sent to the analytical laboratory at the test site menal GmbH. The remaining samples were stored as retain samples at ≤ -18 °C until finalization of the study. Detailed information of the analytical method is given in the analytical phase report.

Preparation of unknown samples at menal GmbH: Samples were immediately prepared either after sample drawing. Samples were filtered through a 0.2 μm regenerated cellulose filter (15 mm) directly into the HPLC vial. The first drops were discarded.
Vehicle:
no
Details on test solutions:
PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution was prepared by adding 35.0 – 35.1 mg test item to 7 L test medium and stirring for ca. 2 h using a magnetic stirrer at room temperature.
The direct addition of the test item into the test vessel filled with test medium in combination with stirring using an adequately dimensioned magnetic stirrer and stir bar was decided to be a more efficient an accurate method than using an overhead shaker with screw-cap glass bottles. This does not impact the quality and reliability of the study negatively.
Test organisms (species):
Danio rerio (previous name: Brachydanio rerio)
Details on test organisms:
TEST ORGANISM
- Common name: Danio rerio
- Source: in-house breeding of Hydrotox GmbH
- Age at study initiation (mean and range, SD): app. 5 months
- Length at study initiation (length definition, mean, range and SD): app. 2.5 cm
- Method of breeding/ Maintenance of the brood fish: The zebrafish are cultured at 26 ± 2 °C with 14 h light : 10 h dark and aeration with air pumps. They are kept in glass tanks, equipped with external filters, at ≤ 1 fish/L. At least every two weeks, ≥ ⅓ of the tank volume is replaced with fresh charcoal filtered tap water.

Ten days before the start of the test, the fish were kept under test conditions in test medium. They were fed every working day with commercial fish food (TetraMin) until 24 h before test start, when feeding was stopped.
- Type and amount of food during acclimation: commercial fish food (TetraMin flake food, Tetra GmbH, Melle, Germany) and two times per week with freshly hatched Artemia salina larvae.

FEEDING DURING TEST: No
Test type:
semi-static
Water media type:
freshwater
Limit test:
yes
Total exposure duration:
96 h
Remarks on exposure duration:
renewal of test solutions after 24 h, 48 h and 72 h
Post exposure observation period:
The absence of any mortality indicates that, according to the Threshold Approach for Acute Fish Toxicity, fish is not the most sensitive group of test organisms after short-term exposure and no further testing (i.e. full EC50 Fish Acute Toxicity Test) is required.
Hardness:
Not reported.
Test temperature:
22.2 - 22.9 °C (in the control) and 22.6 - 23.2 °C (in the test item treatment)
pH:
7.7 -7.8 (in the control) and 7.6 - 7.8 (in the test item treatment)
Dissolved oxygen:
8.0 - 8.4 mg/L (in the control) and 8.1 - 8.3 mg/L (in the test item treatment)
Salinity:
Not reported.
Conductivity:
695 - 701 μS/cm (in the control) and 698 - 701 μS/cm (in the test item treatment)
Nominal and measured concentrations:
nominal concentration (mg/L): 5.0
measured concentration (mg/L): 0.80 (geometric mean)
Details on test conditions:
TEST SYSTEM
- Test vessel: Glass tanks
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: 8 L
- Type of flow-through (e.g. peristaltic or proportional diluter): No: Semi-static system
- Renewal rate of test solution (frequency/flow rate): Yes, after 24 h
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
The test was performed with ISO 7346 medium according to EN ISO 7346 (1997). After addition of 50 mL DF1 and DF2 as well as 120 mL DF3 and DF4, the medium was filled up to 50 L, continuously aerated.
- Source/preparation of dilution water:
- Total organic carbon:
- Particulate matter:
- Metals:
- Pesticides:
- Chlorine:
- Alkalinity:
- Ca/mg ratio:
- Culture medium different from test medium:
- Intervals of water quality measurement:

OTHER TEST CONDITIONS
- Adjustment of pH: 7.7 – 7.8 in the control and 7.6 – 7.8 in the test item treatment
- Photoperiod: 14 h light : 10 h dark
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : mortality and vitality of the fish was assessed after 24 h, 48 h, 72 h and 96 h exposure

TEST CONCENTRATIONS
- Spacing factor for test concentrations: no, only one concentration of the test item was exposed.to test organism
- Justification for using less concentrations than requested by guideline:
- Range finding study: An Algal Growth Inhibition Test (Report No. 1165, see also section 6.1.5) and a Daphnia magna Acute Immobilisation Test (Report No. 1166, see also section 6.1.3) was performed before start of this study. The applyied nominal concentration is based on the measured EC50 from the Algal Growth Inhibition Test
- Test concentrations: 5 mg/L (nominal concentration), 0.80 mg/L (geometric mean concentration)
- Results used to determine the conditions for the definitive study: The lowest EC50 from these two tests result from the Algal Growth Inhibition Test (analytically measured geometric mean test item concentration => threshold concentration):
• Growth Rate: EC50 = 0.37 mg/L
• Yield: EC50 = 0.22 mg/L
Reference substance (positive control):
no
Key result
Duration:
96 h
Dose descriptor:
LOEC
Effect conc.:
> 0.8 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Key result
Duration:
96 h
Dose descriptor:
NOEC
Effect conc.:
>= 0.8 mg/L
Nominal / measured:
meas. (geom. mean)
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
- Behavioural abnormalities: No obvious effect on appearance and behaviour of the fish in the test item treatment was observable. All test organisms in the control as well as in the test item treatment appeared healthy and lively throughout the exposure.
Reported statistics and error estimates:
Determination of propyl gallate in fish growth medium - Phase report:
Calibration curve was back calculated by quadratic regression with 1/y^2 weighting. Final data were generated by the MassHunter 4.0 software and transferred to MS Excel® 2010 for preparation of result tables to present in the final phase report.
Validity criteria fulfilled:
yes
Conclusions:
As no fish died until the end of the exposure, the mortality in the control as well as in the test item treatment was 0 %. As no effect on the test organisms was observable, no statistical data evaluation was performed. As the measured test item concentrations are not within ± 20 % of the nominal concentrations, according to OECD 203 (1992) and OECD 23 (2000), all results are given in relation to the analytically measured test item concentrations. Therefore, the geometric mean of all eight measurements (0 h, 24 h old/new, 48 old/new, 72 h old/new and 96 h) was calculated. For measurements which are < LOQ, the value of LOQ, was used for calculation:
LOEC: ≤ 0.80 mg/L; NOEC: < 0.80 mg/L.
Executive summary:

In a 96-h acute toxicity study, zebra fish (Danio rerio) were exposed to propyl gallate at nominal concentrations of 0.0 (control) and 5 mg/L (measured concentrations of 0.00 (control) and 0.80 mg/L) under semi-static conditions. The NOEC value, based on mortality and sublethal effects, was ≥ 0.80 mg/L, respectively.  Sublethal effects were not observed. Based on the results of this study, propyl gallate would be not classified as toxic to zebra fish in accordance with the CLP classification system.

This toxicity study is classified as acceptable and satisfies the guideline requirement for the zebra fish acute toxicity study.

 

Results Synopsis

 

Test organism size/age: Danio rerio (ca. 2.5 cm, ca. 5-month-old)

Test Type: semi-static

 

NOEC:  ≥ 0.80 mg/L                     

LOEC: ≥ 0.80 mg/L

Endpoint(s) Effected: mortality and abnormal effects

Description of key information

In a 96-h acute toxicity study, zebra fish (Danio rerio) were exposed to propyl gallate at nominal concentrations of 0.0 (control) and 5 mg/L (measured concentrations of 0.00 (control) and 0.80 mg/L) under semi-static conditions (limit test). The NOEC value, based on mortality and sublethal effects, was ≥ 0.80 mg/L, respectively.  The LOEC was > 0.80 mg/L. Sublethal effects were not observed.

Key value for chemical safety assessment

Additional information